Tricyclic derivatives of indole with antiangiogenic activity

ABSTRACT

Compounds of formula 
                 
 
useful for treating tumors, particularly mammary carcinoma and colon carcinoma.

The invention described herein relates to compounds having tricyclicstructure of the type tetrahydrociclopent[b]indole (1),tetrahydrocarbazole (2), and esahydrocycloept[b]indole (3), a processesfor their preparations, and pharmaceutical composition containing thesame for treating tumor and diseases associated with abnormalangiogenesis.

Said compounds have the following general formula (I):

wherein:

X=CH, N

X₁=O, S, N, CH

R and R₁, which may be the same or different, are selected from thegroup consisting of: —H, OH, OR₅ in which R₅ may be C₁-C₄ alkyl orbenzyl, when two groups OR₅ are vicinal R₅ is methylene; or R and R₁ maybe independently nitro; amino possibly mono- or di-substituted withC₁-C₄ alkyl; carboxy; alkoxy (C₁-C₄) carbonyl;

R and R₁ taken together may form an aliphatic or aromatic cyclic grouphaving 5 or 6 atoms;

when X₁=N, CH, then

R₂ is selected from the group consisting of —H, phenyl, benzyl, linearor branched C₁-C₆ alkyl;

n=is an integer ranging from 0 and 4;

R₃, which may be the same as or different from R₄, may be: —H, —OH,—OR₆, wherein

R₆ is linear or branched C₁-C₄ alkyl, or when R₃=R₄=OR₆ vicinal, R₆ isisopropylidene

R₇=C₁-C₄ linear or branched alkyl possibly substituted with one or twogroups OH, OR₆, in case of 2 groups OR₆ vicinal, R₆ is isopropyliden; orR₇ is formyl (CHO), oxime (CH═NOH).

The invention includes all the possible isomers, stereoisomers and theirmixtures, metabolites and their metabolic precursors or bio-precursors(so called pro-drug) of the general formula.

The use of antineoplastic drugs in human therapy causes a substantialnumber of toxic or side effects which consequently lead to a reductionof the amount of drug to be administered, and in some cases todiscontinuation of the therapy. A reduction of the amount of drug to beadministered or discontinuation of the therapy cause an increase inprimary tumour growth and/or the occurrence of tumour metastases.

The growth of a primary tumour is favoured by good vascularisation (byangiogenesis) of the tumour tissue. An adequate supply of oxygen andnutrients promotes rapid growth of the tumour itself. It has beendemonstrated that the extent of angiogenesis can be an extremelynegative factor in the prognosis of neoplasms.

Angiogenesis in the adult is normally quiescent, but it represents anormal function, for example in the healing of wounds, or in thereconstruction of the endometrium during the female reproductive cycle.

The angiogenic response is physiologically stimulated when the vascularfunctions are reduced and tissue perfusion is inadequate.

More generally, it can be claimed that, in physiological conditions,angiogenesis constitutes a positive feedback in response to inadequateperfusion, or to a reduced supply of oxygen and nutrients, such asoccurs, for instance, in the case of occlusion of an artery, insituations of tissue mass growth (for example, the neovascularisationthat accompanies the formation of muscle tissue); and in the case of anincreased work load in association with an increased oxygen and nutrientrequirement.

In the course of local ischaemia, due to partial or complete occlusionof an artery, the development of collateral vessels is necessary inorder to maintain perfusion.

As above mentioned, it has been demonstrated that the extent ofangiogenesis can be an extremely negative factor in the prognosis ofneoplasms (van Hinsbergh V W, Collen A, Koolwijk P; Ann. Oncol., 10Suppl., 4:60-3, 1999; Buolamwini J K; Curr. Opin. Chem. Biol.,3(4):500-9, 1999 August).

It is also known, in the neoplastic field, that a fundamental stage inthe biology of the tumour cell is the acquisition of metastasisingcapability.

The tumour cells that metastasise are able to lose adherence to thesurrounding structures, invade blood and lymphatic vessels and coloniseother tissues at a distance where they can continue to reproducethemselves.

Metastasising is also a critical event in the clinical history of thedisease, being the main cause of death due to cancer. It is closedassociated with and facilitated by the presence of vascular tissue inthe tumour site or adjacent areas.

The migration of tumour cells across the surrounding structures enablesthe cells to reach the intratumoural blood vessels, whether pre-existingor formed by neo-angiogenesis, and thus reach the bloodstream (Ray J M.,Stetler-Stevenson W G; Eur. Respir. J., 7(11):2062-72, 1994;Stetler-Stevenson W G, Liotta L A, Kleiner D E Jr.; FASEB J.,7(15):1434-41, 1993 December).

The presence of communication between lymphatic and blood vessels in thevascular region of the tumour enables the neoplastic cells to move inboth vascular systems.

Recent studies have shown a direct relationship between angiogenesis andarthritic disease (Koch A E; Arthritis and Rheumatism 41:951-962, 1998).In particular, it has been demonstrated that neo-vascularisation of thearticular cartilages plays a crucial role in pannus formation and inprogression of arthritis. A normal cartilage does not possess bloodvessels, while the synovial fluid of arthritic patients contains anangiogenesis-stimulating factor produced by endothelial cells (EASF).

The presence of this factor is associated with vascularisation anddegradation of the cartilage.

Other diseases are also related to abnormal angiogenesis.

It has been found that, in diabetic retinopathy [Histol. Histopathol.1999 October; 14(4):1287-94], psoriasis [Br J. Dermatol. 1999 December;141(6):1054-60], chronic inflammation and arteriosclerosis [Planta Med.1998 December; 64(8):686-95], neovascularisation of the affected tissuesis a facilitating factor.

Over the past thirty years compounds with a cycloalkanoindole structurehave been synthesised and studied with a view to exploiting theirpossible therapeutic potential.

The basic requisite of these compounds—the substituted indole inposition 3—is a feature shared by natural products such as melatonin ortryptophan.

In the '70s, cycloalkanoindole compounds were studied for theirantiinflammatory properties (J. Med. Chem. 1976, 19(6):787-92) or fortheir antidepressant properties (J. Med. Chem. 1976, 19(6):792-7).

These studies were then followed by others (on a number ofaminotetrahydrocarbazols) to assess their effects on the CNS (J. Med.Chem. 1977, 20(4):487-92) in that they possessed a tryptamine-likestructure.

In the '80s, a number of derivatives with a tetrahydrocarbazol structurewere found to possess antibacterial properties: in culture they inhibitthe growth of Trypanosoma cruzi (Rev. Argent. Microbiol. 1987,19(3):121-4).

In the '90s, compounds with a cycloalkanoindole structure were studiedas potential analgesics (Xenobiotica 1989, 19(9):991-1002), with effectson the serotonin receptors (J. Med. Chem. 1993, 36(13): 1918-9) and onthe melatonin receptors (Eur. J. Pharmacol. 1995, 287(3):239-43).

In the past few years, tetrahydrocarbazol derivatives have been studiedfor their antiproliferative properties (Farmaco 1998, 53(6):431-7); inparticular, the N-pyridinium derivative may act with a mechanisminvolving inhibition of topoisomerase II.

U.S. Pat. No. 5,017,593 describes derivatives of cyclohept[b]indolealkanoic acid as leukotriene antagonists.

EP 0496237 describes N-imidazolyl derivatives of tetrahydrocarbazol andcyclohept[b]indoles as thromboxane antagonists (TXA-2), useful in thetreatment of cardiovascular disorders (myocardial infarction andangina), cerebrovascular disease (stroke, transient ischaemic attacks,migraine), peripheral vascular disease (microangiopathy), kidney disease(glomerular sclerosis, lupus nephritis, diabetic nephropathy),respiratory disease (bronchoconstriction and asthma) andatherosclerosis.

J. Med. Chem. 1998, 41, 451-67 describes compounds withtetrahydrocyclopent[b]indole (1), tetrahydrocarbazol (2), andhexahydrocyclohept[b]indole (3) structures used for studies on melatoninreceptors having formula:

(1) n = 0 tetrahydrocyclopent[b]indole (2) n = 1 tetrahydrocarbazole (3)n = 2 hexahydrocycloept[b]indole

U.S. Pat. No. 5,830,911; U.S. Pat. No. 4,927,842; U.S. Pat. No.4,616,028 describe tetrahydrocarbazol compounds, with antiinflammatoryactivity having formula:

R = CN; R′ = H R = F, NO₂; R′ = CN R = R′ = CN COX-2 inhibitorAntiarthritic U.S. Pat. No. 5830911

R = CH₂CN; CH₂COCH₃; R = CH₂CH═CH₂ CH₂CH═N—OCH₃; AY-30068 CH₂CH═N—OH;(CH₂)₄—CH═CH₂ U.S. Pat. No. 4927842 U.S. Pat. No. 4616028Antiinflammation Antiinflammation and antiangiogenic

The compounds described in the above cited publications are differentfrom those claimed in the present invention.

Despite the progress made in recent years, the pharmacological researchconcerned with discovering new drugs for the treatment of tumor diseasesand diseases characterised by abnormal angiogenesis is still consideredby many experts in medicine as one of the most promising field.

In fact, to date there is still a strongly perceived need for newcompounds capable of blocking or interfering with the tumour diseasesand diseases caused by abnormal angiogenesis. As mentioned above, thesediseases include tumours, tumours metastasis, arthritic diseases,diabetic retinopathy, psoriasis, chronic inflammation andarteriosclerosis.

It has now been found that the formula (I) compounds, characterised bythe presence of two aromatic bases (indole or one of its derivatives),where the first is condensed to a saturated cycle in position 2-3, ofthe tetrahydrocarbazol type, and the second aromatic base, bound inposition 3, is present as a substituent in the benzyl position of thesaturated ring, unexpectedly possess antitumor and antiangiogenicproperties.

Compounds with general formula (I) are therefore the object of theinvention described herein.

A further object of the invention described herein are compounds withgeneral formula (I) and their use in the medical field.

A further object of the invention described herein are compounds withgeneral formula (I) and a process for their preparation.

A further object of the invention described herein is a pharmaceuticalcomposition containing as active ingredient a formula (I) compound andat least a pharmaceutically acceptable excipient and/or diluent.

A further object of the invention described herein is a pharmaceuticalcomposition containing as active ingredient a formula (I) compound, forthe treatment of a tumour pathology, in which the tumour is selectedfrom the group consisting of sarcoma, carcinoma, carcinoid, bone tumour,neuroendocrine tumour, lymphoid leukaemia, acute promyelocyticleukaemia, myeloid leulaemia, monocytic leukaemia, megakaryoblasticleukaemia and Hodgkin's disease.

A further object of the invention described herein is the use ofcompounds of formula (I) for the preparation of a medicament withantiangiogenic activity.

A further object of the invention described herein is the use ofcompounds of formula (I) for preventing the onset of tumour metastases.

A further object of the invention described herein is the use ofcompounds of formula (I) for the treatment of arthritic disease.

A further object of the invention described herein is the use ofcompounds of formula (I) for the treatment of diabetic retinopathy.

A further object of the invention described herein is the use ofcompounds of formula (I) for the treatment of psoriasis.

A further object of the invention described herein is the use ofcompounds of formula (I) for the treatment of chronic inflammatorydiseases.

A further object of the invention described herein is the use ofcompounds of formula (I) for the treatment of arteriosclerosis.

As mentioned above, the growth of a primary tumour is facilitated bygood vascularisation of the tumour tissue, and the extent of theneoangiogenesis may be a highly adverse factor in the prognosis ofneoplasms. An adequate supply of oxygen and nutrients in the tumoursite, in fact, facilitates rapid growth of the tumour itself.

It is well known that the therapeutic measures available to physiciansfor the treatment of tumours are still unable to prevent many patientsfrom dying of these diseases. It is also well known that mostoncological patients are treated not with a single anticancer drug butwith a combination of several anticancer agents. The need to administeranticancer drugs in combination stems from the fact that by acting atdifferent metabolic levels in some cases they favour complete remissionof the tumour, while in others they lengthen the patient's life and/orimprove the quality of life of the patients treated.

To date there is still a strongly perceived need for new compounds to beused in combination with known compounds in the fight against cancer.

The compound according to the invention described herein can be used incombination with one or more anticancer drugs.

A further object of the invention described herein is the combination ofcompounds of formula (I) with one or more known anticancer drugs.

A further object of the invention described herein is a pharmaceuticalcomposition containing the combination of compounds of formula (I) withone or more known anticancer drugs, and one or more excipients orvehicles pharmacologically acceptable.

A further object of the invention described herein is a pharmaceuticalcomposition containing as active ingredient a formula (I) compound, incombination with one or more known antitumour compounds, in which theantitumour compound is selected from the group consisting of alkylatingagents, topoisomerase inhibitors, antitubulin agents, intercalatingcompounds, anti-metabolites, natural products such as vinca alkaloids,epipodophyllotoxins, antibiotics, enzymes, taxans, andcyto-differentiating compounds.

A further object of the invention described herein is the use of thecombination of compounds of formula (I) and the anticancer compound toprepare a medicament for the treatment of tumour, characterised in thatthe compound of formula (I) is present as a coadjuvant of the anticancercompound.

The following examples illustrate the invention

The synthesis of cyclised products consists of two stages: the firststage consists in the condensation, in the geminal position, of ahydroxyaldehyde with 2 aromatic bases; the second stage consists in acyclisation reaction with DAST (diethyl amino-sulphur trifluoride). Thissynthetic sequence, albeit with exceptions, may represent the synthesisprocess adopted in the preparation of all the derivatives describedherein. For the sake of simplicity of description, the case of formula(I) compounds where dove X is CH, X₁ is NH, and R₃ and R₄ are hydrogenis illustrated. It is perfectly clear that the expert in the sector canprepare all the formula (I) compounds using suitable starting materialsand adopting suitable reagents, simply by availing himself or herself ofhis own general knowledge, or with the aid of the standard manualsavailable.

As the expert in the sector will readily appreciate, the first stage inthe synthesis involves the preparation of intermediate products with abisindole structure.

Their preparation can be done using various different methods.

PROCEDURE A: Synthesis of Derivatives with Mannofuranose

Reaction (Tetrahedron Asymmetry, 1997, 8(17), 2905-12): The indole orits derivatives (1 mmol) was dissolved in Et₂O anhydrous (50 ml). Asolution of ethylmagnesiumbromide/ether (3M) (0.33 ml; 1 mmol) wasslowly added. The solution so obtained was left under stirring, inanhydrous conditions, for several minutes: a magnesium white saltderivative was obtained. The ether was evaporated the white residualobtained was dissolved in anhydrous CH₂Cl₂. The solution was left atroom temperature/reflux for 12/36 h.

Work-up: the solution was quenched by addition of a saturated solutionof NaHCO₃/10% NH₄Cl. The organic phase was separated and dried on Na₂SO₄and evaporated. The desired product was purified by flash chromatography(hexane/acetone).

PROCEDURE B: Synthesis of Hydroxyaldeide Derivatives

Reaction: the indole or its derivative (2 mmol) was dissolved with thealdehyde (5-hydroxy-pentanale or 2-etoxytetrahydrofurano) (1 mmol), in15 ml of MeOH/H₂O (2/1). finally Dysprosium triflate was added and themixture was left to react at room temperature/80° C. for 6/36 h.

Work-up: The reaction mixture was quenched with 10% NaHCO₃, extractedwith CH₂Cl₂. The extracted were dried Na₂SO₄, and evaporated. The rawresidual was purified by preparative-HPLC and two regioisomers wereisolated (X) and (Y).

Procedure C: Reaction of Cyclization on Mannofuranose Derivatives

Reaction: The bis-indolyl derivative (476 mg; 1 mmol) was dissolved inCH₂Cl₂ (80 ml). At the solution was added, at room temperature,diethylaminosulfur trifluoride (DAST) (400 μl; 3 mmol). The reaction wasrapid.

Work-up: after 60′ a solution of 10% NaHCO₃ was added, and the solutionwas extracted with CH₂Cl₂. The organic extracted were dried on Na₂SO₄and evaporated. The reaction products present in this raw reactionproduct were isolated by preparative-TLC or better by preparative-HPLCRP-18.

Procedure D: Reaction of Cyclization on Derivatives with Hydroxyaldehyde

Reaction: the symmetric derivative (1) or asymmetric derivative (2) (1mmol) was dissolved in CH₂Cl₂ (20 ml). DAST (200 μl; 1.5 mmol) was addedto the solution, at 0° C.—room temperature. The reaction was rapid.After 15′-20′ the starting product was almost completely reacted.

Work-up: after 30′ a solution of 10% NaHCO₃ was added, the solution soobtained was extracted with CH₂Cl₂. The organic extracted were dried onNa₂SO₄ and evaporated. The reaction products present in this rawreaction product were isolated by preparative-TLC or better by RP-18prep.-HPLC.

Starting from the symmetric or from the non-symmetric product wereformed both, the derivative with the second indole residual toward thelower part (3) with a yield of 30-60% and the derivative with the indoleresidual toward the high part (4): the latter is present in 10-25%respect to the other.

Procedure E: Debenzylation Reaction

Reaction: the benzylated derivative (1 mmol) was dissolved in CH₃OH (50ml)

The catalyst (10% Pd/C; 30 mg) was added to the solution, at roomtemperature.

The solution so obtained was left under hydrogen (60 psi). After 16 hthe starting product was completely reacted.

Work-up: The catalyst was filtered of. The organic phase was evaporated.The deprotected product was purified by flash-chromatography. Yield 85%.

Procedure F: Deprotection Reaction

Reaction: the protect product (1 mmol) was dissolved in tetrahydrofuran(THF) (50 ml). HCl 1N was added to the solution. The solution soobtained was left for 1 h at 20° C.-40° C. So the starting product wascompletely reacted. The principal deprotect product obtained was thedesired product.

With reference to the deprotected product only in the esocyclicresidual, the deprotection can be obtained by acid hydrolysis at lowtemperature (ex. 1N HC at low temperature, for 30′-60′).

Work-up: the obtained product (vedi es. 1) was shacked with a saturatedsolution of NaHCO₃. The THF was evaporated, then the product wasextracted with AcOEt. The organic phase was evaporated end thedeprotected product was purified by flash-chromatography.

Yield 85%.

Procedure G: Oxydation Reaction.

Reaction: The aldehyde (1 mmol) was dissolved in 20 mL of MeOH. To thesolution was added NaIO4 (1 mmol) dissolved in 2 mL of H₂O.

The solution so obtained was left at room temperature for 4 h.

Work-up: 1° Step: a Na₂S₂O₃ aqueous solution was added. The organicsolvent was evaporated and the residual was extracted with AcOEt.

2° Step: the protect intermediate can be deprotected as described inscheme 6.

EXAMPLE 1

ST 1345

(R,S)-5-hydroxy-1,1-(indol-2-yl,indol-3-yl)-pentan

TLC (Exane/Isopropanol=97.5/2.5): 0.5 HPLC RP-18 Waters 250×4.6 (70%H₂O, 30% CH₃CN, Flow 1 ml/min): 6.16 NMR 300 MHz (H-1, CDCl₃): H₁(4.30t, 1H)-H₅(3.5 t, 2H)-H₂₋₄(1.4-1.5-2.1 m, 6H)-H_(1′b)-H_(1′a) (7.8-8.0 s,2H)-H_(4′b)(7.5 m, 1H)-H_(7′a)(7.4 d, 1H)-H_(7′b)(7.22 d,1H)-H4′a-_(5′a-5′b-6′a-6′b-)(6.9-7.2 m, 5H)-H_(2′a)(6.96 d,1H)-H_(3′b)(6.4 d, 1H) Ion Spray (M⁻): 317 Elemental analysis:(calculated) C, 79.21%; H, 6.96%; N, 8.79%. (found) C, 78.64%; H, 7.15%;N, 8.45%.

EXAMPLE 2

ST 1346

5-Hydroxy-1,1-di-(indol-3-yl)-pentane

TLC (Exane/iPrOH=97.5/2.5): 0.43 HPLC RP-18 Waters 250×4.6 (40% H₂O, 60%CH₃CN, Flow 1 ml/min): 5.36 NMR 300 MHz (H-1, CDCl₃): H₁(4.7 t,1H)-H₅(3.8 t, 2H)-H₄(2.3 m, 2H)-H₃(1.7 m, 2H)-H₂(1.8 m, 2H)-H_(2′)(7.2s, 2H)-H_(5′)(7.15 t, 2H)-H_(6′)(7.4 t, 2H)-H_(4′)(7.5 d, 2H)-H_(7′)(7.8d, 2H)-H_(1′)(8.15 br.s, 2H) NMR 300 MHz (C-13, CDCl₃):C₁(34.2)-C₂(33.0)-C₃(24.6)-C₄(35.7)-C₅(63.2)-C_(7′)(111.2)-C_(6′)(119.2)-C_(4′)(119.8)-C_(3′)(120.4)-C_(5′)(121.6)-C_(2′)(122)-C_(3′bis)(127.2)-C_(7′bis)(136.7)Ion Spray (M⁻): 317 Elemental analysis: C, 79.21%; H, 6.96%; N, 8.79%.found C, 78.70%; H, 7.29%; N, 8.39%. Melting point: 190° C. (dec)

EXAMPLE 3

ST 1422

5-hydroxy-1,1-di-(5,6-methylendioxy-indol-3-yl)-pentane

TLC (Hexane/iPrOH=97.5/2.5): 0.55 HPLC RP-18 (50% H₂O, 50% CH₃CN, Flow 1ml/min): 6.7 NMR 300 MHz (H-1, CD₃CN): H₁(4.2 t, 1H)-H₂(2.2 m,2H)-H₃-H₄(1.4-1.5 m, 2H)-H₅(3.4 q, 2H)-H_(8′)(5.8 s,4H)-H_(4′)-H_(7′)(6.8 s, 4H)-H_(2′)(7.0 s, 2H)-H_(1′)(8.9, brs, 2H) IonSpray (M⁺): 407 Elemental analysis: C, 67.97%; H, 5.46%; N, 6.89%. foundin accordance with the theoretical. Melting point: 200° C. (dec.)

EXAMPLE 4

ST 1423

(R,S)-5-hydroxy-1,1-di-(5′,6′-methylendioxy-indol-2-yl,5″,6″-methylendioxy-indol-3-yl)-pentane

TLC (Hexane/iPrOH=97.5/2.5): 0.63 HPLC RP-18 (50% H₂O, 50% CH₃CN, Flow 1ml/min): 8.2 NMR 300 MHz (H-1, CD₃CN): H₁(4.3 t, 1H)-H₂(2.2 m, 2H)-H₃-H₄(1.4-1.6 m, 4H)-H₅(3.5 m, 2H)-H_(8′a)-_(8′b) (5.9 s, 4H)-H_(3′b)(6.3 s,1H)-H_(4′a)-H_(7′a) (6.8-6.9 d, 2H)-H_(4′b)-H_(7′b) (7.0 d,2H)-H_(2′a)(7.2 s, 1H)-H_(1′a-1′b)(8.9-9.1 brs-brs, 2H). NMR 300 MHz(C-13, CD₃CN):25.1-33.4-34.9-37.2-62.5-92.6-93.0-98.4-99.2-99.7-101.3-101.5-121.7 IonSpray (M⁺): 407 Elemental analysis: (calculated) C, 67.97%; H, 5.46%; N,6.89%. (found) in accordance with the theoretical. Melting point: 220°C. (dec.)

EXAMPLE 5

ST1730

4-hydroxy-1,1-di-(5′,6′-methylendioxy-indol-3-yl)-butane

TLC (Hexane/iPrOH=75/25): 0.44 HPLC RP-18 (60% H₂O, 40% CH₃CN, Flow 1ml/min): 17.5 NMR 300 MHz (H-1, CD₃CN): H₁(4.3 t, 1H)-H₂(2.3 m,2H)-H₃(1.6 m, 2H)-H₄(3.6 q, 2H)-H_(8′)(6.0 s, 4H)-H_(4′)-H_(7′)(6.9 s,4H)-H_(2′)(7.2 s, 2H)-H_(1′)(9.0 brs, 2H) Ion Spray (M⁺): 393 Elementalanalysis: C, 67.34%; H, 5.14%; N, 7.14%. (found): in accordance with thetheoretical. Melting point: 240° C. (dec.)

EXAMPLE 6

ST1731

(R,S)-4-hydroxy-1,1-di-(5′,6′-methylendioxy-indol-2-yl,5″,6″-methylendioxy-indol-3-yl)-butane

TLC (Hexane/iPrOH=75/25): 0.41 HPLC RP-18 (60% H₂O, 40% CH₃CN, Flow 1ml/min): 23.4 NMR 300 MHz (H-1, CD₃CN): H₁(4.3 t, 1H)-H₂(2.2 m,2H)-H₃(1.6 m, 4H)-H₄(3.7 m, 2H)-H_(8′a)-_(8′b) (6.0 s, 4H)-H_(3′b)(6.4s, 1H)-H_(4′a)-H_(7′a) (6.8-6.9 d, 2H)-H_(4′b)-H_(7′b) (7.0 d,2H)-H_(2′a)(7.2 s, 1H)-H_(1′a-1′b)(8.9-9.1 brs-brs, 2H) NMR 300 MHz(C-13, CD₃CN):29.8-30.4-35.5-60.8-90.9-91.4-96.7-97.5-98.1-99.6-99.9-116.6-116.9-119.9-121.6-130.0-130.9-141.3-141.5-141.6-142.9-143.8Ion Spray (M⁻): 391 Elemental analysis: (calculated): C, 67.34%; H,5.14%; N, 7.14%. (found): in accordance with the theoretical. Meltingpoint: 205° C. (dec.)

EXAMPLE 7

ST 1707

1,1-di(indol-3-yl)-4-hydroxy-butane

TLC (Hexane/AcOEt=1/1): 0.26 HPLC RP-18 (50% H₂O, 50% CH₃CN, Flow 1ml/min): 8.3 NMR 300 MHz (H-1, CD₃CN): H₁(4.3 t, 1H)-H₂(2.2 m,2H)-H₃(1.4 m, 2H)-H₄(3.4 t, 2H)-H_(2′)(7.2 s, 2H)-H_(5′)(6.8 t,2H)-H_(6′)(6.9 t, 2H)-H_(4′)(7.2 d, 2H)-H_(7′)(7.5 d, 2H)-H_(1′)(10.7br.s, 2H) NMR 300 MHz (C-13, CD₃CN):C₁(61.5)-C₂₋₃(32.2)-C₄(34)-C_(7′)(112)-C_(6′)(118.5)-C_(3′)-C_(4′)(119.5-119.7)-C_(5′)(121.2)-C_(2′)(121.6)-C_(3′bis)(127.4)-C_(7′bis)(137.1)Ion Spray (M⁻): 303 Elemental analysis: (calculated) C, 78.92%; H,6.62%; N, 9.20%. (found) in accordance with the theoretical. Meltingpoint: 110-115° C.

EXAMPLE 8

ST 1750

4-hydroxy-1,1-di-(5′,6′-methylendioxy-indol-2-yl)-butane

TLC (Hexane/iPrOH=75/25): 0.60 HPLC RP-18 (60% H₂O, 40% CH₃CN, Flow 1ml/min): 19.9 NMR 300 MHz (H-1, CD₃CN): H₁(4.3 t, 1H)-H₂(2.3 m,2H)-H₃(1.6 m, 2H)-H₄(3.6 q, 2H)-H_(8′)(6.0 s, 4H)-H_(4′)-H_(7′)(6.9-7.02s, 4H)-H_(3′)(6.4 s, 2H)-H_(1′)(9.1 brs, 2H) Ion Spray (M⁻): 391Elemental analysis: C, 67.34%; H, 5.14%; N, 7.14%. (found): inaccordance with the theoretical. Melting point: 250° C. (dec.)

EXAMPLE 9

ST 1866

1,1-di(7′-aza-indol-3-yl)-4-butanol

TLC (Hexane/AcOEt=75/25): 0.18 HPLC RP-18 (60% H₂O, 40% CH₃CN, Flow 1ml/min): 4.4 NMR 300 MHz (H-1, CD₃OD): H₁(4.4 t, 1H)-H₂(2.3 m,2H)-H₃(1.6 m, 2H)-H₄(3.6 t, 2H)-H_(2′)(7.3 s, 2H)-H_(5′)(6.9 m,2H)-H_(4′)(8.1 t, 2H)-H_(6′)(7.8 d, 2H) NMR 300 MHz (C-13, CD₃CN):30.3-30.7-33.6-61.0-114.0-117.2-119.4-121.9-127.5-140.9-147.7 Ion Spray(M⁻): 305 Elemental analysis: (calculated) C, 70.57%; H, 5.92%; N,18.29%. (found) in accordance with the theoretical. Melting point: 221°C. (dec.)

EXAMPLE 10

ST 1372

1-(indol-3-yl)-2,3-O-isopropylidene-4-(2,3-O-isopropylidene-ethyl)-tetrahydrocarbazole

TLC (Hexane/Acetone=8/2): 0.75 HPLC RP-18 Waters 300×3.3(40% H₂O, 60%CH₃CN, Flow 1 ml/min): 12.8

Racemic Mixture

NMR 300 MHz (H-1, CH₃CN): H₁+H₃ (4.52 m, 2H)-H₂(4.64 m, 1H)-H₄(3.66 m,1H)-H₅(4.67 m, 1H)-H₆(3.97/3.69 m, 2H)-H₈(1.42 s, 3H)-H₉(1.32 s,3H)-H₁₁(1.46 s, 3H)-H₁₂(1.38 s, 3H)-H_(2′a)(6.94 s, 1H)-H_(4′a)(7.51 d,1H)-H_(4′b)(7.69 d, 1H)-H_(6′a)(7.16 m, 1H)-H_(6′b)(7.09 m,1H)-H_(7′a)(7.47 d, 1H)-H_(7′b)(7.26 d, 1H)-H_(1′a)(9.0 s,1H)-H_(1′b)(8.3 s, 1H) NMR 300 MHz (C-13, CH₃CN): C₁(38.15), C₂(80.77),C₃(75.9), C₄(40.77), C₅(78.0), C₆(68.33), C₈(25.5), C₉(26.8), C₁₁(25.9),C₁₂(28.3), C_(2′a)(124.5), C_(2′b)(135.7), C_(3′a)(115.9),C_(3′b)(107.6), C_(6′a)(122.7), C_(6′b)(121.9), C_(7′a)(112.5),C_(7′b)(111.9), C_(8′a)(127.5), C_(8′b)(128.2), C_(9′b)(137.7),C_(9′a)(137.9). Ion Spray (M⁺) 459 Elemental analysis: (calculated) C,73.34%; H, 6.59%; N, 6.11%. (found) C, 73.11%; H, 6.63%; N, 5.55%.Melting point: 204-206° C. (dec.)

EXAMPLE 11

ST 1381

1-(indol-3-yl)-indo[2,3a]-cycloheptan.

TLC (Hexan/iPrOH=95/5): 0.22 HPLC RP-18 Waters 300×3.3 (40% H₂O, 60%CH₃CN, Flow 1 ml/min): 22.1

Racemic Mixture

NMR 300 MHz (H-1, DMSO-d6): H₁(4.65 br.s,1H)-H₂(1.95 e 2.4 mm,2H)-H₃(1.7 m, 2H)-H₄(1.6 e 1.9 mm, 2H)-H₅(2.75 e 3.0 mm,2H)-H_(2′a)(6.68 s, 1H)-H_(5′a)-H_(5′b)(6.95 m, 2H)-H_(6′a)(7.07 t,1H)-H_(4′b)(7.45 m, 1H)-H_(7′b)(7.2 m, 1H)-H_(7′a)(7.37 d,1H)-H_(4′a)(7.52 d, 1H)-H_(1′a)(10.8 s,1H)-H_(1′b)(10.4 s, 1H). NMR 300MHz (C-13, DMSO-d6): C₁(36.3), C₂(33.2), C₃(26.5), C₄(28.6), C₅(24.0),C_(7′b)(110.5), C_(7′a)(111.4), C_(3′a)(115.6), C_(4′b)(117.2),C_(5′b)(117.7), C_(5′a)(118.2), C_(4′a)(118.5), C_(6′b)(119.7),C_(6′a)(120.7), C_(2′a)(123.5), C_(8′a)(125.9), C_(8′b)(128.6),C_(9′b)(134.2), C_(9′a)(136.6), C_(2′b)(139.7). Ion Spray (M⁻): 299Elemental analysis: (calculated) C, 83.96%; H, 6.71%; N, 9.33%. (found)C, 81.19%; H, 6.50%; N, 9.03% Melting point: 206-208° C.

EXAMPLE 12

ST 1621

5-(indol-3-yl)-indo[2,3a]-cycloheptan.

TLC (Hexan/iPrOH=95/5): 0.15 HPLC RP-18 (40% H₂O, 60% CH₃CN, Flow 1ml/min): 16.6

Racemic Mixture

NMR 300 MHz (H-1, DMSO-d6): H₅(4.8 br.s,1H)-H₄(1.9 and 2.4 mm,2H)-H₃(1.5 and 1.7 mm, 2H)-H₂(1.5 and 1.9 mm, 2H)-H₁(2.9 br.s,2H)-H_(2′a)(6.5 s, 1H)-H_(5′b)(6.8 t, 1H)-H_(6′b)(6.92 t,1H)-H_(5′a)(6.97 t, 1H)-H_(6′a)(7.07 t, 1H)-H_(4′b)(7.13 d,1H)-H_(7′b)(7.24 d, 1H)-H_(7′a)(7.3 d, 1H)-H_(4′a)(7.62 d,1H)-H_(1′a)(10.6 s,1H)-H_(1′b)(10.7 s, 1H). NMR 300 MHz (C-13, DMSO-d6):C₃(25.2)-C₂(27.3)-C₁(28.3)-C₅(31.8)-C₄(33.7)-C_(7′b)(10.2)-C_(7′a)(111.3)-C_(3′b)(114.5)-C_(4′b)(117.1)-C_(3′a)(117.4)-C_(5′b)(117.7)-C_(5′a)(117.9)-C_(4′a)(118.6)-C_(6′b)(119.6)-C_(6′a)(120.5)-C_(2′a)(123.5)-C_(8′a)(126.2)-C_(8′b)(128.5)-C_(9′b)(134.3)-C_(9′a)(136.6)-C_(2′b)(137.4).Ion Spray (M⁻): 299 Elemental analysis: (calculated) C, 83.96%; H,6.71%; N, 9.33%, in accordance with the theoretical. Melting point: 170°C.

EXAMPLE 13

ST 1728

1-(1H-indol-3-yl)-tetrahydro-1H-carbazole

TLC (Hexane/iPrOH=9/1): 0.66 HPLC RP-18 (40% H₂O, 60% CH₃CN, Flow 1ml/min): 18.4

Racemic Mixture

NMR 300 MHz (H-1, CD₃CN): H₁(4.46 t, 1H)-H₂(2.1 and 2.3 mm, 2H)-H₃(1.9and 2.1 mm, 2H)-H₄(2.81 t, 2H)-H_(2′a)(6.98 s, 1H)-H_(5′a)(6.92 t,1H)-H_(5′b)-H_(6′b) (7.0 m, 2H)-H_(6′a)(7.09 m, 1H)-H_(7′b)(7.15 m,1H)-H_(4′a)(7.29 d, 1H)-H_(7′a)(7.4 dt, 1H)-H_(4′b)(7.46 d,1H)-H_(1′a)(9.1 br.s, 1H)-H_(1′b)(8.6 br.s, 1H). NMR 300 MHz (C-13,,CD₃CN):C₄(21.8)-C₃(23.1)-C₂(33.0)-C₁(33.2)-C_(3′b)(110.6)-C_(7′b)(111.4)-C_(7′a)(112.3)-C_(4′b)(118.5)-C_(3′a)(118.7)-C_(5′b)(119.4)-C_(5′a)(119.7)-C_(4′a)(119.8)-C_(6′b)(121.5)-C_(6′a)(122.4)-C_(2′a)(123.9)-C_(8′a)(127.5)-C_(8′b)(128.6)-C_(9′b)(137.0)-C_(9′a)(137.8)-C_(2′b)(137.9).Ion Spray (M⁺): 287 Elemental analysis: (calculated) C, 83.30%; H,6.99%; N, 9.71%, in accordance with the theoretical. Melting point: 207°C.

EXAMPLE 14

ST 1729

4-(1H-indol-3-yl)-tetrahydro-1H-carbazole

TLC (Hexane/iPrOH=9/1): 0.55 HPLC RP-18 (40% H₂O, 60% CH₃CN, Flow 1ml/min): 12.4

Racemic Mixture

NMR 300 MHz (H-1, CD₃CN): H₁(2.83 m, 1H)-H₂(1.95 and 1.84 mm,2H)-H₃(2.17 and 2.04 mm, 2H)-H₄(4.49 t, 1H)-H_(2′a)(6.77 d,1H)-H_(4′a)(7.43 d, 1H)-H_(6′a)(7.06 t, 1H)-H_(7′a)(7.36 d,1H)-H_(5′a)(6.92 m, 1H)-H_(7′b)(7.27 d, 1H)-H_(4′b)(6.83 d, 1H)-H_(5′a)(6.71 t, 1H)-H_(6′b)(6.94 m, 1H)-H_(1′a)(8.96 br, 1H)-H_(1′b)(8.94 br,1H). NMR 300 MHz (C-13, CD₃CN):C₂(22.0)-C₁(24.0)-C₄(31.0)-C₃(33.0)-C_(7′b)(111.3)-C_(7′a)(112.3)-C_(3′b)(112.5)-C_(4′a)(119.3)-C_(3′a)(120.9)-C_(6′b)(121.2)-C_(6′a)(122.1)-C_(2′a)(123.7)-C_(8′a)(127.7)-C_(8′b)(128.5)-C_(2′b)(136.2)-C_(9′b)(137.0)-C_(9′a)(137.8).Ion Spray (M⁺): 287 Elemental analysis: (calculated) C, 83.30%; H,6.99%; N, 9.71%. Melting point: 182° C.

EXAMPLE 15

ST 1749

1-(5″,6″-methylendioxy-indol-3-yl)-5′,6′-methyleridioxy-indo[2,3-a]-cycloeptane

TLC (Hexane/AcOEt=8/2): 0.23 HPLC RP-18 (40% H₂O, 60% CH₃CN, Flow 1ml/min): 15.4

Racemic Mixture

NMR 300 MHz (H-1, CD₃CN): H₁(4.45 m, 1H)-H₂(2.05 and 2.25 mm, 2H)-H₃-H₄(1.8 m, 4H)-H₅(2.85 m, 2H)-H_(10′b)(5.85 d, 1H)-H_(10′a)(5.90 s,2H)-H_(7′b)(6.66 s, 1H)-H_(2′a)(6.8 d, 1H)-H_(4′b)(8.3 s,1H)-H_(4′a)(6.9 s, 1H)-H_(7′a)(6.94 s, 1H)-H_(1′b)(8.4 s,1H)-H_(1′a)(9.0 s,1H). NMR 300 MHz (C-13, CD₃CN): C₅(25.4), C₃-C₄(29.5),C₂(35.3), C₁(38.1), C_(7′b)(92.4), C_(7′a)(93.0), C_(4′a)(97.3),C_(4′b)(98.4), C_(10′b)(101.1), C_(10′a)(101.5), C_(3′b)(113.4),C_(3′a)(118.2), C_(8′a)(121.3), C_(2′a)(121.7), C_(8′b)(124.0),C_(9′b)(129.8), C_(9′a)(132.7), C_(2′b)(139.3), C_(6′b)(142.9),C_(6′a)(143.3), C_(5′b)(144.3), C_(5′a)(145.5). Ion Spray (M⁺):389Elemental analysis: (calculated) C, 71.12%; H, 5.19%; N, 7.21%. (found)in accordance with the theoretical. Melting point: 184° C. (dec.)

EXAMPLE 16

ST 1751

1-(5′,6′-methylendioxy-1H-indol-3-yl)-6,7-methylendioxy-tetrahydro-1H-carbazole

TLC (Hexane/iPrOH=9/1): 0.31 HPLC RP-18 (40% H₂O, 60% CH₃CN, Flow 1ml/min): 11.5

Racemic Mixture

NMR 300 MHz (H-1, CD₃CN): H₁(4.32 t, 1H)-H₂(2.2 and 2.0 mm, 2H)-H₃(2.03and 1.83 mm, 2H)-H₄(4.3 t, 2H)-H_(10′a)-H_(10′b)(5.8-5.9 dd,4H)-H_(4′a)(6.65 s, 1H)-H_(7′b)(6.72 s, 1H)-H_(2′a)(6.88 d,1H)-H_(7′a)-H_(4′b)(6.90 s, 1H)-H_(1′b)(8.4 s, 1H)-H_(1′a)(9.0 s, 1H).NMR 300 MHz (C-13, CD₃CN):C₄(21.8)-C₃(23.2)-C₂(33.0)-C₁(33.3)-C_(7′b)(92.8)-C_(7′a)(93.0)-C_(4′b)(97.6)-C_(4′a)(98.2)-C_(10′a)(101.2)-C_(10′b)(101.5)-C_(3′b)(110.8)-C_(3′a)(119.1)-C_(8′a)(121.4)-C_(2′a)-C_(8′b)(122.4)-C_(9′b)(131.6)-C_(9′a)(132.6)-C_(2′b)(136.6)-C_(6′b)(142.9)-C_(6′a)(143.2)-C_(5′b)(144.6)-C_(5′a)(145.4).Ion Spray (M⁺): 375 Elemental analysis: (calculated) C, 70.58%; H,4.85%; N, 7.48%, in accordance with the theoretical. Melting point: 200°C. (dec.)

EXAMPLE 17

ST 1765

1-(5″-benzyloxy-indol-3-yl)-5′-benzyloxy-indo[2,3a]-cycloheptan.

TLC (Hexan/iPrOH=9/1): 0.62 HPLC RP-18 (40% H₂O, 60% CH₃CN, Flow 1ml/min): 27.1

Racemic Mixture

NMR 300 MHz (H-1, CD₃CN): H₁(4.54 d, 1H)-H₂(2.32 and 2.08 mm,2H)-H₃(1.83 m, 2H)-H₄(1.83 m, 2H)-H₅(2.88 m, 2H)-H_(10′a)(4.99 s,2H)-H_(10′b)(5.13 s, 2H)-H_(5′a)-H_(5′b)(6.95 m, 2H)-H_(6′b)(6.74 q,1H)-H_(6′a)(6.87 q, 1H)-H_(2′a)(6.90 d, 1H)-H_(4′a)(6.93 d,1H)-H_(7′b)(7.07 d, 1H)-H_(4′b)(7.09 d, 1H)-H_(14′a)-H_(14′b)(7.31-7.27m, 2H)-H_(7′a)(7.34 m, 1H)-H_(13′a)(7.33 m, 1H)-H_(13′b)(7.38 m,1H)-H_(12′a)(7.39 d, 1H)-H_(12′b)(7.47 d, 1H)-H_(1′b)(8.47s,1H)-H_(1′a)(9.0 s, 1H). NMR 300 MHz (C-13, CD₃CN): C₁(38.01),C₂(35.1), C₃(29.5), C₄(29.4), C₅(25.3), C_(10′a)(71.3) C_(10′b)(71.4),C_(4′b)(102.5), C_(4′a)(103.7), C_(6′b)(111.7), C_(7′b)(112.0),C_(7′a)(113.1), C_(3′b)-C_(6′a) (113.2), C_(3′a)(117.4), C_(2′a)(125.0),C_(8′a)(127.9), C_(12′b)(128.5), C_(12′a)(128.6), C_(13′a)(129.3),C_(13′b)(129.4), C_(8′b)(130.5), C_(9′b)(130.7), C_(9′a)(133.2),C_(11′a)(139.0), C_(11′b)(139.3), C_(2′b)(141.7), C_(7′a)(113.0),C_(6′a-)C_(3′a) (113.2), C_(14′a-)C_(14′b-)C_(12′b) (128.5),C_(12′a)(128.6) Ion Spray (M⁺): 513 Elemental analysis: (calculated) C,82.00%; H, 6.29%; N, 5.46%.—(found) in accordance with the theoretical.Melting point: 286° C. (dec.)

EXAMPLE 18

ST 1777

4-(5′,6′-methylendioxy-1H-indol-3-yl)-6,7-methylendioxy-tetrahydro-1H-carbazole

TLC (Hexane/iPrOH=9/1): 0.20 HPLC RP-18 (40% H₂O, 60% CH₃CN, Flow 1ml/min): 7.9

Racemic Mixture

NMR 300 MHz (H-1, CD₃CN): H₁(2.80, 1H)-H₂(1.96 and 1.84 mm, 2H)-H₃(2.13and 1. mm, 2H)-H₄(4.31 t, 1H)-H_(10′b)(5.76 d, 2H)-H_(10′a)(5.86 d,2H)-H_(4′b)(6.22 s, 1H)-H_(4′a)(6.74 s, 1H)-H_(2′a)(6.75 d,1H)-H_(7′b)(6.82 s, 1H)-H_(7′a)(6.88 s, 1H)-H_(1′b)(8.83 s,1H)-H_(1′a)(8.86 s, 1H). NMR 300 MHz (C-13, CD₃CN):C₂(22.3)-C₁(23.9)-C₄(31.7)-C₃(33.1)-C_(7′b)(92.7)-C_(7′a)(93.0)-C_(4′b)(98.3)-C_(4′a)(98.5)-C_(10′b)(101.1)-C_(10′a)(101.4)-C_(3′b)(112.8)-C_(3′a)(120.9)-C_(8′a)(121.5)-C_(8′b)-C_(2′b)(122.3)-C_(9′b)(131.6)-C_(9′a)(132.6)-C_(2′b)(134.7)-C_(6′b)(142.4)-C_(6′a)(142.9)-C_(5′b)(144.2)-C_(5′a)(145.2).Ion Spray (M⁻): 373 Elemental analysis: (Calculated) C, 70.20%; H,5.36%; N, 7.44%, in accordance with the theoretical. Melting point: 228°C. (dec.)

EXAMPLE 19

ST 1778

5-(5″-benzyloxy-indol-3-yl)-5′-benzyloxy-indo[2,3a]-cycloheptan.

TLC (Hexane/iPrOH=9/1): 0.4 HPLC RP-18 (40% H₂O, 60% CH₃CN, Flow 1ml/min): 18.1

Racemic Mixture

NMR 300 MHz (H-1, CD₃CN): H₁(2.89 m,1H)-H₄(2.49 m, 2H)-H₃(1.6 and 1.8mm, 2H)-H₂(1.6 m, 2H)-H₅(4.72 t, 1H)-H_(10′b)(4.89 d, 2H)-H_(10′a)(5.08s, 2H)-H_(2′a)(6.54 d, 1H)-H_(6′b)(6.68 q, 1H)-H_(4′b)(6.75 d,1H)-H_(6′a)(6.83 q, 1H)-H_(7′b)(7.16 d, 1H)-H_(4′a)(7.16 d,1H)-H_(7′a)(7.28 d, 1H)-H_(13′b)(7.29 m, 1H)-H_(12′b)(7.34m,1H)-H_(13′a)(7.37 m, 1H)-H_(12′a)(7.47 d, 1H)-H_(1′a)(8.90 s,1H)-H_(1′b)(8.93 s, 1H). NMR 300 MHz (C-13, CD₃CN):C₃(26.6)-C₂(28.5)-C₁(29.7)-C₅(33.4)-C₄(34.7)-C_(10′b)(71.2)-C_(10′a)(71.4)-C_(4′b)(112.5)-C_(4′a)(104.0)-C_(6′b)(111.6)-C_(6′a)(111.8)-C_(7′b)(112.9)-C_(7′a)(112.9)-C_(3′b)(116.1)-C_(3′a)(119.2)-C_(2′a)(125.3)-C_(8′a)(128.1)-C_(14′b)(128.5)-C_(14′a)(128.6)-C_(12′b)(128.6)-C_(12′a)(128.7)-C_(13′a)(129.2)-C_(13′b)(129.3)-C_(8′b)(130.5)-C_(9′b)(131.0)-C_(9′a)(133.3)-C_(11′a)-C_(11′b)(139.1)-C_(2′b)(139.7)-C_(5′a)(153.3)-C_(5′b)(153.5).Ion Spray (M⁻): 511 Elemental analysis: (calculated) C, 82.00%; H,6.29%; N, 5.46%, in accordance with the theoretical. Melting point: 237°C. (dec.)

EXAMPLE 20

ST1783

1-(5″-hydroxy-1H-indol-3-yl)-5′-hydroxy-indo[2,3a]-cycloheptan.

TLC (Hexane/iPrOH=9/1): 0.35 HPLC RP-18 Waters 300×3.3 (45% H₂O, 55%CH₃CN, Flow 1 ml/min): 4.2

Racemic Mixture

NMR 300 MHz (H-1, CD₃CN): H₁(4.61 d, 1H)-H₂(2.3 and 2.0 mm, 2H)-H₃(1.9m, 2H)-H₄(1.8 m, 2H)-H₅(2.97 m, 2H)-OH(6.45 br.d, 2H)-H_(7′b)(6.67 d,1H)-H_(7′a)(6.83 d, 1H)-H_(4′a) H_(4′b)(6.95 s, 2H)-H_(2′a)(7.0 s,1H)-H_(6′b)(7.1 d, 1H)-H_(6′a)(6.4 d, 1H)-H_(1′b)(8.44s,1H)-H_(1′a)(9.01 s, 1H). NMR 300 MHz (C-13, CD₃CN): C₁(36.6),C₂(33.6), C₃(28.1), C₄(28.0), C₅(23.8), C_(4′b)(101.1), C_(4′a)(102.4),C_(6′b)(109.1), C_(7′b)(110.1), C_(7′a)(110.7), C_(3′b)-C_(6′a) (111.0),C_(3′a)(115.3), C_(2′a)(126.5), C_(8′a)(128.4), C_(8′b)(128.6),C_(9′b)(129.2), C_(9′a)(131.0), C_(2′b)(140.1), C_(5′a-)C_(5′b)(149.3).Ion Spray (M⁺): 333 Elemental analysis: (calculated) C, 75.88%; H,6.06%; N, 8.43%. (found) in accordance with the theoretical.

EXAMPLE 21

In the similar way were prepared the following compounds:

COMPOUND PROCEDURE NOTE ST1866 A Bis-7-azaindolyl via magnesiumbromideST1345 B Bis-indolyl via acid catalysis ST1346 B ″ ST1422 B ″ ST1423 B ″ST1707 B ″ ST1750 B ″ ST1372 C Cyclization of sugar moieties ST1381 DCyclization of the non-sugar moieties ST1621 D ″ ST1728 D ″ ST1729 D ″ST1749 D ″ ST1751 D ″ ST1765 D ″ ST1777 D ″ ST1778 D ″ ST1783 EDebenzylation ST1900 F Deprotection ST1901 F Deprotection ST (all) GOxydation

Pharmacology

The abbreviation ST, followed by a number, identifies the compoundsfiguring in the examples reported in the pharmacological assays.

For the antiangiogenic activity the chemotactic assay with the Boydenchamber was performed (Werner F., Goodwin R. H. and Leonard E. J.,Journal of Immunological Methods 1980; 33, 239-247), using both bovineaortal endothelial cell (BAEC) cultures and bovine medullary endothelialcell (BMEC) cultures. The assays were performed at IC₀ (maximumnon-cytotoxic concentration) and the results expressed as % inhibitionof migration across a porous filter in response to a chemotacticstimulus (1% bovine serum in DMEM culture medium). This finding wasobtained by direct cell count under the optical microscope, and thepercentage migration inhibition was calculated according to the formula(T−C/C)×100, where T=mean number of cells migrating in the sample andC=mean number of cells migrating in the control. The control consistedof cells which migrated towards the serum, not treated with the studymolecules, and included in every chemotaxis experiment. The data referto the readings of 5 microscopic fields/well in 4 independent chemotaxiswells per sample. The results obtained are reported on Table 4.

For the cytotoxic activity, proliferation screening tests were used,with different tumour lines, such as MCF-7 (human mammary carcinoma),LoVo (human colon carcinoma), MES-SA (human uterine sarcoma), or K-562(human chronic myeloid leukaemia).

The test used was the sulforodamine B test used for screening anticancerproducts at the National Cancer Institute (Skehan, 1990). The moleculesat scalar concentrations over a range from 500 μM to 0.97 μM, wereincubated in parallel with different human cell lines for 24 hours.After removing the products, cell survival was investigated afteranother 48 hours with the NCI test. The antiproliferative capacity ofthe compounds was quantified in terms of IC₅₀±SD (concentration of themolecule that inhibits 50% of cell survival) processed using a curvefitting program (De Lean et al., 1978). The results obtained arereported on Table 1 and Table 2.

The cell cycle and apoptosis analysis on a tumour line was done byincubating the products for 24 hours at a concentration equal toapproximately the IC₅₀ values with MCF-7 cells. The molecules wereremoved and the cell cycle and apoptosis were assessed at differenttimes (0, 24, 48 hours). The cells were stained with propidium iodideand analysed with a cytofluorimeter (FACS) (Beckman Dickinsonfluorescence activated cell sorter) by means of an argon ion laser setat 488 nm for the excitation. To assess the percentage of cells in thevarious stages of the cycle, linear DNA histograms were analysed with acell fit program distributed by the equipment manufacturer. For theanalysis of apoptosis, a region was inserted below the G0/G1 peak of thecontrol population and the data were analysed with software supplied bythe company (Lysis II-C32). The results obtained are reported on Table3.

The cytotoxicity of the molecules on tumour lines resistant tochemosensitising activity was assessed on various tumour cell linesoverexpressing P-glycoprotein and resistant to doxorubicin (100-fold)and cross-resistant to daunorubicin, actinomycin D, mitoxantrone,vincristine, vinblastine, taxol, colchicine, and etoposide. Thecytotoxicity of the products was assessed using the same test adoptedfor the sensitive tumour cells.

Later, the products were assayed at a concentration lower than or equalto the one that inhibits 10% of cell survival. At this concentration,the molecules were tested in parallel in the absence and presence ofdoxorubicin. MDR ratios were calculated for the IC₅₀ values in order toestablish the degree of potentiation of the cytotoxic activity ofdoxorubicin induced by the product (MDR ratio) (De Lean et al. (1978) A.J. Physiol. 235, E97-102); (Skehan et al. (1990) J. Natl. Cancer Inst.82, 1107-1112).

TABLE 1 ANTIPROLIFERATIVE ACTIVITY vs SENSITIVE CELL IC₅₀ +/− SD (μM)Series Compound MCF-7 LoVo Tetrahydrocarbazoles ST1372 21.1 +/− 0.3 21.7 +/− 4.3  ″ ST1728  22 +/− 2.8 22.8 +/− 2.9  ″ ST1729  35 +/− 6.150.5 +/− 4.8  ″ ST1777 0.96 +/− 0.19  0.64 +/− 0.003Esahydrocycloept[b]indoles ST1381 28.5 +/− 1.8  22.5 +/− 2.6  ″ ST162122.5 +/− 1.3  34.8 +/− 4.3  ″ ST1765 11.5 +/− 1.1  10.5 +/− 0.07 ″ST1778 28.5 +/− 1.4  54.5 +/− 7.4  ″ ST1783 27 +/− 4    29 +/− 0.06

TABLE 2 ANTIPROLIFERATIVE ACTIVITY vs RESISTENT CELL IC₅₀ +/− SD (μM)Series Compound MCF-7/DX LoVo/DX Tetrahydrocarbazoles ST1372 26.9 +/−1.6  26.3 +/− 3.7  ″ ST1751 58.6 +/− 9   EsahydroCycloept[b]indolesST1381 28.4 +/− 3   25.2 +/− 4.2  ″ ST1621 68.3 +/− 4.6  23.8 +/− 3.7  ″ST1765 24.8 +/− 3.8  17.03 +/− 0.91  ″ ST1778  71 +/− 0.4 65.1 +/− 8.1 ″ ST1783   39 +/− 0.03

TABLE 3 CELLULAR CYCLE AND APOPTOSIS ON MCF-7 CELL Apoptosis CompoundG0/G1 (%) S (%) G2 + M (%) 48 h (%) ST1372 C = 43.7 C = 47.6 C = 8.6 C =3.3 40 μM = 57.8 40 μM = 36.8 40 μM = 5.4 40 μM = 16 20 μM = 54.4 20 μM= 34.2 20 μM = 11.4 ST1381 C = 43.8 C = 47.6 C = 8.6 C = 3.3 30 μM =57.7 30 μM = 32.2 30 μM = 10.1 30 μM = 3.3

ST1372 at 40 μM increases and blocks in G0/G1 32% of the cell, and at 20μM increases and blocks in G0/G1 23% of the cell.

ST1372 at 11 μM increases 3.5 times the activity of the Doxorubicin bothon MCF-7/Dx line and on LoVo/Dx cell line.

ST1381 at 30 μM increases and blocked in G0/G1 32% of the cell line; isnot cytotoxic vs endotelial cell (IC>100 μM); and is active for thechemotaxys.

TABLE 4 CYTOTOXYCITY AND CHEMOTAXYS ON BMEC MEC % inhibition IC₅₀ IC₀ ofmigration Series Compound (μM) (μM) at IC₀/D.S.Esahydrocycloept[b]indoles ST1381 >100 30 −61.7 +/− 6.3 ″ ST1621 10 0.1−43 +/− 4 ″ ST1749 >200 100 −40 +/− 3 ″ ST1778 50 25 −40 +/− 4 ″ ST178360 10 −46 ″ ST1729 80 25 −40 +/− 3

Although ST 1381 results the better anti-chemiotactic compound, isimportant to note that all compounds of this group decrease thechemiotaxis of endothelial cells.

The composition according to the invention contain as active ingredientat least one formula (I) alone or in combination with other activeingredients useful in the treatment of the diseases indicated in theinvention described herein, in the form of separate doses or in formssuitable for combined therapies. The active ingredient according to theinvention will be in a mixture with appropriate vehicles and/orexcipients commonly used in pharmacy, such as, for instance, thosedescribed in “Remington's Pharmaceutical Sciences Handbook”, latestedition. The compositions according to the invention will contain atherapeutically effective amount of the active ingredient. The doseswill be determined by the expert in the sector, for example theclinician or primary-care physician, according to the type of disease tobe treated and the patient's condition, or concomitantly in conjunctionwith the administration of other active ingredients.

Examples of pharmaceutical compositions are those that allow oral orparenteral, intravenous, intramuscular, subcutaneous or transdermaladministration. Pharmaceutical compositions suitable for the purpose aretablets, rigid or soft capsules, powders, solutions, suspensions,syrups, and solid forms for extempore liquid preparations. Compositionsfor parenteral administration are, for example, all the intramuscular,intravenous, and subcutaneous injectable forms, in the form ofsolutions, suspensions or emulsions. Also worthy of mention are theliposomal formulations. Suitable compositions also include forms basedon slow release of the active ingredient, whether as oral administrationforms, tablets coated with suitable layers, microencapsulated powders,cyclodextrine complexes, or depot forms. e.g. subcutaneous, such asdepot injections or implants.

1. A compound having the formula (I):

wherein: X=CH X₁=N R and R₁, which are the same or different, areselected from the group consisting of —H, OH, OR₅ in which R₅ may beC₁-C₄ alkyl or benzyl, when two groups OR₅ are vicinal R₅ is methylene;or R and R₁ may be independently nitro; amino optionally mono- ordi-substituted with C₁-C₄ alkyl; carboxy; or alkoxy (C₁-C₄) carbonyl; Rand R₁ taken together form an aliphatic or aromatic cyclic group having5 or 6 atoms; R₂ is selected from the group consisting of —H, phenyl,benzyl, linear or branched C₁-C₆ alkyl; n=is an integer ranging from 0and 4; R₃, which is the same as or different from R₄, is —H, —OH, —OR₆,wherein R₆ is linear or branched C₁-C₄ alkyl, or when R₃=R₄=OR₆ vicinal,R₆ is isopropylidene

R₇=C₁-C₄ linear or branched alkyl optionally substituted with one or twogroups OH, OR₆, in case of 2 groups OR₆ vicinal, R₆ is isopropylidene;or R₇ is formyl (CHO), oxime (CH═NOH), their isomers and their mixtures.2. A pharmaceutical composition containing as active ingredient acompound of claim 1, and at least a pharmaceutically acceptableexcipient and/or diluent.
 3. A combination consisting of (a) a compoundof formula (I) as claimed in claim 1 with (b) an anticancer drug, inwhich the anticancer drug is selected from the group consisting ofalkylating agents, topoisomerase inhibitors, antitubulin agents,intercalating compounds, anti-metabolites, natural products,epipodophyllotoxins, antibiotics, enzymes, taxans, cyto-differentiatingcompounds and anti-angiogenic compounds.
 4. A pharmaceutical compositioncomprising as active ingredient the combination of claim 3 and one ormore pharmacologically acceptable excipients or vehicles.
 5. Thecomposition of claim 4, wherein the compound of formula (I) is presentas a co-adjuvant of the anticancer compound.
 6. The composition of claim4, wherein the compound of formula (I) and the anticancer drugs areadministered simultaneously or sequentially.
 7. The composition of claim2 or 4, in the form of tablets, capsules, powders, solutions,suspensions, vials, syrups, suppository, enema, foam or liposomalformulations, useful for oral, parenteral or rectal administration.
 8. Amethod of treating mammary carcinoma or colon carcinoma comprisingadministering to a subject in need thereof a therapeutically effectiveamount of a compound of claim 1.